Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a biophysical technique well suited to the characterization of protein dynamics and protein-ligand interactions. In order to accurately define the rate of exchange, HDX experiments require the repeated measure of deuterium incorporation into the target protein across a range of time points. Accordingly, the HDX-MS experiment is well suited to automation, and a number of automated systems for HDX-MS have been developed. The most widely utilized platforms all operate an integrated design, where robotic liquid handling is interfaced directly with a mass spectrometer. With integrated designs, the exchange samples are prepared and injected into the LC-MS following a "real-time" serial workflow. Here we describe a new HDX-MS platform that is comprised of two complementary pieces of automation that disconnect the sample preparation from the LC-MS analysis. For preparation, a plate-based automation system is used to prepare samples in parallel, followed by immediate freezing and storage. A second piece of automation has been constructed to perform the thawing and LC-MS analysis of frozen samples in a serial mode and has been optimized to maximize the duty cycle of the mass spectrometer. The decoupled configuration described here reduces experiment time, significantly improves capacity, and improves the flexibility of the platform when compared with a fully integrated system.

中文翻译：

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用于氢/氘交换质谱实验的解耦自动化平台。

氢/氘交换质谱 (HDX-MS) 是一种生物物理技术，非常适合表征蛋白质动力学和蛋白质-配体相互作用。为了准确定义交换率，HDX 实验需要在一系列时间点内重复测量氘掺入目标蛋白的情况。因此，HDX-MS 实验非常适合自动化，并且已经开发了许多用于 HDX-MS 的自动化系统。使用最广泛的平台都采用集成设计，其中机器人液体处理直接与质谱仪连接。通过集成设计，交换样品按照“实时”串行工作流程制备并注入 LC-MS。在这里，我们描述了一个新的 HDX-MS 平台，它由两个互补的自动化部分组成，将样品制备与 LC-MS 分析分开。在制备过程中，使用基于板的自动化系统并行制备样品，然后立即冷冻和储存。已经构建了第二个自动化部分，以在串行模式下对冷冻样品进行解冻和 LC-MS 分析，并进行了优化，以最大限度地提高质谱仪的占空比。与完全集成的系统相比，此处描述的解耦配置减少了实验时间，显着提高了容量，并提高了平台的灵活性。然后立即冷冻和储存。已经构建了第二个自动化部分，以在串行模式下对冷冻样品进行解冻和 LC-MS 分析，并进行了优化，以最大限度地提高质谱仪的占空比。与完全集成的系统相比，此处描述的解耦配置减少了实验时间，显着提高了容量，并提高了平台的灵活性。然后立即冷冻和储存。已经构建了第二个自动化部分，以在串行模式下对冷冻样品进行解冻和 LC-MS 分析，并进行了优化，以最大限度地提高质谱仪的占空比。与完全集成的系统相比，此处描述的解耦配置减少了实验时间，显着提高了容量，并提高了平台的灵活性。